Biolayer Interferometry (BLI) is a powerful technique that enables direct measurement of biomolecular interactions in real time without the need for labeled reagents. Here we describe the analysis of a high-affinity binding interaction between a monoclonal antibody and purified antigen using BLI. A simple Dip-and-Read™ format in which biosensors are dipped into microplate wells containing purified or complex samples provides a highly parallel, user-friendly technique to study molecular interactions. A rapid rise in publications citing the use of BLI technology in a wide range of applications, from biopharmaceutical discovery to infectious diseases monitoring, suggests broad utility of this technology in the life sciences.